Histopathological findings in the early diagnosis of granulosa cell tumour in bitches

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call‐Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call‐Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call‐Exner body.     

Effect of cumulus cell removal and sperm pre‐incubation with progesterone on in vitro fertilization of equine gametes in the presence of oviductal fluid or cells

In spite of many attempts to establish an in vitro fertilization (IVF) technique in the equine, no efficient conventional IVF technique is available. The presence of oviductal fluid or oviductal cells during IVF helps to improve embryo production in vitro but is not sufficient to reach high fertilization rates. Thus, our aim was to perform equine IVF either after sperm pre‐incubation with oviductal fluid or in the presence of oviductal cells, and to evaluate the effect of cumulus removal from the oocyte or sperm pre‐incubation with progesterone. In experiments 1 and 2, IVF was performed in the presence of porcine oviduct epithelial cells. The removal of cumulus cells from equine oocytes after in vitro maturation tended to increase the percentage of fertilization when fresh sperm was used (1/33 vs. 4/31, p > 0.05) but had no effect when frozen sperm was used (1/32 vs. 1/32). Equine sperm pre‐incubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/14 vs. 2/18 for fresh, 1/29 vs. 1/25 for frozen). In experiments 3 and 4, IVF was performed after pre‐incubation of sperm with porcine oviductal fluid. The removal of cumulus cells tended to increase the percentage of fertilization when fresh sperm was used (1/24 vs. 3/26, p > 0.05). Sperm pre‐incubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/39 vs. 2/36 for fresh, 2/37 vs. 1/46 for frozen), but two 3–4 cell stage zygotes were obtained with fresh sperm pre‐incubated with progesterone. This is an encouraging result for the setting up of an efficient IVF procedure in equine.     

In vitro susceptibility to antimonials and amphotericin B of Leishmania infantum strains isolated from dogs in a region lacking drug selection pressure

The aim of this study was to evaluate the susceptibility to anti-leishmanial agents of 24 strains isolated from dogs living in the urban area of Alger lacking drug selection pressure. Two different Leishmania infantum zymodemes, MON-1 and MON-281, were identified in these dogs. The in vitro susceptibility to the main forms of antimonial and amphotericin were assessed on promastigote and amastigote life stages in culture. The results obtained for both parasite life stages were concordant whatever the molecule tested. Moreover, our data showed that isolates belonging to the relatively rare zymodeme of L. infantum, MON-281, were less susceptible to antimony than MON-1, when at the same time there was no significant difference for amphotericin B.     

Evaluation of the on-line electrical conductivity of milk in mastitic dairy cows

Mastitis is a persistent, inflammatory reaction of the udder tissue, which entails a decline in potassium, and is also responsible for a higher somatic cell count (SCC) and electrical conductivity (EC) of milk. The measurement of EC is an indirect, rapid method to detect subclinical mastitis from milk. The aim of this study was to analyse the EC of milk throughout the lactation, around the day of mastitis detection, and also to estimate its heritability based on data from a large-scale dairy farm. Shortly after calving the EC value generally decreases; however, it was discovered that from the thirteenth week onwards, substantial differences arise between the mastitic and healthy groups of cows. The authors observed a significant (P < 0.001) increase in EC before the detection of clinical mastitis. This higher value (around 11 mS) persisted for 4 days, then it gradually returned to the normal level. The EC of milk from daughters sired by different bulls responded differently in case of mastitis. The heritability of EC seems to be high (h2 = 0.56). Therefore, the EC trait can be a beneficial indicator in detecting mastitis and should be considered in sire selection.     

Flash electroretinography in standing horses using the DTL microfiber electrode

Purpose The goal of our study was the evaluation of a practical method for the recording of flash electroretinograms (ERGs) in sedated, standing horses with the DTL? microfiber electrode. Methods The horses were sedated intravenously with detomidine hydrochloride (0.015 mg/kg). The pupil was dilated and the auriculopalpebral nerve was blocked. The ERGs were recorded with the active electrode on the cornea (DTL?), the reference electrode near the lateral canthus, and the ground electrode over the occipital bone. The light intensities of the white strobe light were 0.03 cd·s/m² (scotopic) and 3 cd·s/m² (scotopic and photopic). Photopic and scotopic single flash and flicker responses to Ganzfeld stimulation were recorded. During the 20‐min dark adaptation period the retina was stimulated every 5 min with the 0.03 cd·s/m² single flash. Results The median b‐wave amplitudes and implicit times were 38 µV and 33 ms (photopic cone‐dominated response), 43 µV and 63 ms (5‐min dark adaptation), 72 µV and 89 ms (10 min), 147 µV and 103 ms (15 min), 188 µV and 109 ms (20 min, 0.03 cd·s/m², rod response), and 186 µV and 77 ms (20 min, 3 cd·s/m², maximal combined rod‐cone response). A steady increase in amplitude and implicit time was noted during dark adaptation. No oscillatory potentials could be isolated. Conclusions The use of detomidine hydrochloride sedation and the DTL? microfiber electrode allowed the recording of good quality ERGs. This protocol should permit the detection of functional problems in the retina without the risk involved with general anesthesia.     

The presence of methicillin-resistant Staphylococcus aureus on large pig breeding farms in Croatia

Methicillin-resistant Staphylococcus aureus (MRSA) have emerged worldwide and have become resistant to a variety of antibiotics. MRSA colonisation in pigs was first reported from the Netherlands in 2005, where pigs were implicated as a source of human MRSA infections (Voss et al., 2005). This paper presents the first report on the presence of MRSA on large pig breeding farms in Croatia, together with the determination of the mecA gene, the results of spa typing and susceptibility to commonly used antimicrobials. Dust samples (7-11 per farm) were collected from eight large pig farms in Croatia. Of the total 68 swabs, the mecA gene was detected in 24 isolates growing on the MRSA agar. All isolates were resistant to oxacillin, tetracycline and streptomycin, and susceptible only to vancomycin, while 92% of the strains were susceptible to ciprofloxacin. Genotyping of the MRSA strains was performed by spa typing, and revealed t011 (n = 17), t034 (n = 5) and t1451 (n = 2). The results presented here predict that MRSA is present on a large number of pig farms in Croatia.     

Effect of diet composition on postweaning colibacillosis in piglets

The weaning of piglets is often associated with digestive disorders, particularly diarrhea--postweaning colibacillosis (PWC)--which is caused by infection with enterotoxigenic strains of Escherichia coli. It has been shown previously that a diet for newly weaned pigs based on cooked white rice and animal protein decreases the occurrence of PWC, whereas the addition of carboxymethylcellulose (CMC) to this diet enhances PWC. The aims of the current work were to 1) determine whether substitution of animal protein with plant proteins in the cooked-white-rice diet influenced its protective effects on PWC and 2) confirm that an increase in viscosity of the digesta by adding CMC to the diet favors the development of PWC--with (Exp. 1) or without (Exp. 2) experimental infection of piglets with E. coli. The diets were 1) cooked white rice and animal protein sources (RAP), 2) RAP + CMC added at 40 g of CMC/kg (air-dry basis) of diet, 3) cooked white rice and plant protein sources (RPP), and 4) wheat and plant protein sources (WPP). Experiments 1 and 2 were conducted using 32 and 24 piglets (eight and six per treatment), respectively. Piglets were weaned at 21 d (d 1), and fed ad libitum until slaughter on d 9. In Exp. 1, piglets were orally infected with enterotoxigenic E. coli on d 4, 5, 6, and 7. On d 8 of Exp. 1, the E. coli scores in feces of pigs fed RAP + CMC were higher than with RAP (P < 0.01). On d 9 after weaning, feces from pigs fed diet RAP were normal or moist, whereas feces from pigs fed RAP + CMC were wet to diarrheic. On d 7 of Exp. 2, pigs fed diets RAP + CMC and WPP had wetter feces than pigs fed diets RAP or RPP (P < 0.05). On d 8, the E. coli scores in feces were higher (P < 0.01) with pigs fed RAP + CMC than with all other diets. The E. coli scores in the digesta were also higher with pigs fed RAP + CMC, and to a lesser extent with diet WPP, than with pigs fed RAP or RPP (P < 0.01). The large intestine was heavier in pigs fed diets RPP and WPP, and the digesta were more acidic (P < 0.05). This study confirmed that diet RAP was protective against PWC, and that substitution of animal proteins with plant protein in a rice-based diet did not diminish its protective effects. The addition of CMC to cooked white rice increased digesta viscosity and enhanced PWC. Consequently, this diet represents a useful model for studying this condition.